摘要
目的探讨亚砷酸钠(NaAsO_(2))对人肝星状细胞(LX-2细胞)活化的影响及机制。方法体外培养LX-2细胞,取对数生长期LX-2细胞,分别加入浓度为0、5、15、20、30、40、50μmol/L的NaAsO_(2)培养液,用MTT法检测细胞活力并确定NaAsO_(2)实验浓度。取对数生长期LX-2细胞随机分为对照组及NaAsO_(2)低、中、高浓度组,分别用0、5、10、15μmol/L的NaAsO_(2)干预48 h。用透射电子显微镜观察细胞自噬变化,用Western blotting法检测细胞活化蛋白(ɑ-SMA)及自噬蛋白(AKT、Hes1、mTOR)表达。结果随着NaAsO_(2)染毒浓度的增加,细胞增殖抑制率逐渐升高;根据细胞半数抑制浓度及细胞活性选择5、10、15μmol/L为NaAsO_(2)实验浓度。对照组可观察到自噬小体、自噬溶酶体。NaAsO_(2)低、中、高浓度组细胞间隙逐渐增宽,观察到“假狄氏间隙”,自噬小体数量增加,脂滴的分布更加密集,可见部分线粒体肿胀、空泡化,大量线粒体嵴断裂;NaAsO_(2)高浓度组以上变化最为明显。与对照组比较,NaAsO_(2)低、中、高浓度组ɑ-SMA蛋白相对表达量逐渐升高(P均<0.05);AKT、mTOR蛋白表达逐渐降低,Hes1蛋白表达逐渐升高(P均<0.05)。结论NaAsO_(2)可诱导LX-2细胞活化,其机制可能与促进细胞自噬有关。
Objective To investigate the effect of sodium arsenite(NaAsO_(2))on activation of human hepatic stellate cells(LX-2 cells)and its mechanism.Methods LX-2 cells were cultured in vitro.The LX-2 cells in the logarithmic growth phase were cultured with NaAsO_(2) medium at the concentrations of 0,5,15,20,30,40,50μmol/L.The cell viability was detected by MTT method and the experimental concentration of NaAsO_(2) was determined.LX-2 cells in the logarithmic growth phase were randomly divided into the control group,low-,medium-and high-concentration NaAsO_(2) groups,which were treated with 0,5,10,and 15μmol/L NaAsO_(2) for 48 h,respectively.The autophagy of the cells was detected by transmission electron microscope,and the expression levels of the cell activated protein(ɑ-SMA)and autophagy protein(AKT,Hes1,and mTOR)were detected by Western blotting.Results With the increase of NaAsO_(2) concentration,the inhibition rate of cell proliferation increased gradually.Five,10,and 15μmol/L were selected as the experimental concentrations of NaAsO_(2) according to the half-maximal inhibitory concentration and the cell viability.Autophagosome and autophagolysosome were observed in the control group.In the low-,medium-and high-concentration NaAsO_(2) groups,the cell gap widened gradually,and"False Dee gap"was observed.The number of autophagosomes increased,and the distribution of lipid droplets was more dense.Some mitochondria were swollen and vacuolated,and a large number of mitochondrial cristae were broken,and the above changes were the most obvious in the high-concentration NaAsO_(2) group.Compared with the control group,the relative expression ofɑ-SMA protein gradually increased,the protein expression of AKT and mTOR gradually decreased,and while the protein expression of Hes1 gradually increased in the low-,medium-and high-concentration NaAsO_(2) groups(all P<0.05).Conclusion NaAsO_(2) may induce the activation of LX-2 cells by promoting autophagy.
作者
李昂
黄菲
迪丽娜尔·亚尔麦麦提
丁关鑫
日沙来提·塔依尔
吴顺华
LI Ang;HUANG Fei;Dilinaer·Yaermaimaiti;DING Guanxin;Rishalaiti·Tayier;WU Shunhua(Department of Occupational and Environmental Health,School of Public Health,Xinjiang Medical University,Urumqi 830011,China;不详)
出处
《山东医药》
CAS
2022年第19期12-15,共4页
Shandong Medical Journal
基金
国家自然科学基金资助项目(82160650,81660523)。
关键词
肝脏疾病
人肝星状细胞
亚砷酸钠
细胞活化
细胞自噬
hepatic disease
human hepatic stellate cells
sodium arsenite
cell activation
cell autophagy
